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Biological assays for irritant, tumor-initiating and -promoting activities

III. Computer-assisted management and validation of biodata generated by standardized initiation/promotion protocols in skin of mice

  • Original Papers
  • Experimental Oncology
  • Published:
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Summary

The initiation/promotion standard protocol 28 (protocol 28), developed and used previously as an experimental model to verify the cancerogenic process of initiation/promotion in mouse skin, was revised in three aspects: (a) statistically it was shown sufficient to use, per promoter dose group, 16 colony-outbred female NMRI mice: (b) by weekly individual records of tumor response (and health status) of each mouse in a dose group, cumulative tumor incidences (and mean and extreme body weights) are determined; from these data the collective records (tumor response, health status), the only data accessible from protocol 28, may be generated in addition; (c) the details of dose groups and all data on tumor response and health status are processed by computer using the program package PAPILLOM. The latter was developed specifically for this purpose, is written in the programming language APL and designed for easy handling by staff of animal laboratories. The program package calculates, from the individual records per promoter dose group, cumulative tumor incidences (and survival data) with confidence limits for any one exposure time, and the package may be linked to programs for statistical validations. In addition, from the collective records it calculates the tumor rates, tumor yields and survival rates for any one exposure time. These data, obtained by either of the standard protocols (16 or 28), are fully comparable. For pure compounds they may be used to calculate semiquantitative tumor-promoting potencies. These values for more than 80 polyfunctional diterpenes of the tigliane, ingenane and daphnane type, scattered in or calculated from previous papers, together with their irritancies, were compiled. Within recent years, computer-assisted standard protocol 16 has been used to handle and evaluate about 1000 promoter dose groups. Protocol 16 allows one to extract and utilize more and better toxicological information on tumor response and health status from any one dose group, utilizing significantly fewer experimental animals than required by protocol 28. Thus, the computer-assisted standard protocol 16 optimizes the utility of the experimental model of mouse skin for the amount, quality and management of experimental data as well as for the requirements of animal protection.

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Abbreviations

δ a :

censoring status of animala

A :

total number of animals

APL:

a programming language

B :

duration of experiment in weeks

DMBA:

7,12-dimethylbenz[a]anthracene

eprf:

environmental principal risk factor

F *(t):

stepwise cumulative incidence of tumors at exposure timet

G ab :

body weight of animala in weekb

LG b :

minimum weight in weekb

M a :

maximum number of tumors of animala (in anyone dose group during promotional period)

MG b :

mean weight in weekb

N ab :

number of individual skin tumors of animala in weekb

rtpp24 :

relative tumor-promoting potency as determined by standard protocols 28 or 16 in female NMRI mice (e.g. at exposure timet=24 weeks)

R T :

tumor response;S, r,b, survival rate in weekb

S *(t):

stepwise cumulative tumor-free survival at exposure timet

TPA:

12-O-tetradecanoylphorbol 13-acetate

T r ,b :

tumor rate in weekb

T y, b :

tumor yield in weekb

T b :

total number of tumors in weekb

TB b :

total number of tumor-bearing animals in weekb

UG b :

maximum weight in weekb

X a :

individual latency time for the first tumor in mousea of any one dose group during promotional period

References

  • Appel KE, Fürstenberger G, Hapke HJ, Hecker E, Hildebrandt AG, Koransky W, Marks F, Neumann HG, Ohnesorge FK, Schulte-Hermann R (1990) Chemical cancerogenesis: definitions of frequently used terms. J Cancer Res Clin Oncol 116:1–5

    Google Scholar 

  • Breslow NE, Edler L, Berger J (1984) A two sample censored-data rank test for acceleration. Biometrics 40:1049–1062

    Google Scholar 

  • Chouroulinkov I, Lasne C, Lowy R, Wahrendorf J, Becher H, Day NE, Yamasaki H (1989) Dose and frequency effect in mouse skin tumor promotion. Cancer Res 49:1964–1969

    Google Scholar 

  • Edler L (1982) Vertrauensbereiche für Quantile bei zensierten Daten — ein Vergleich verschiedener Methoden. EDV in Medizin und Biologie 13:114–119

    Google Scholar 

  • Edler L, Wahrendorf J, Berger J (1980) SURVIVAL — a program package for the statistical analysis of censored survival times. Stat Software Newsl 6:44–53

    Google Scholar 

  • Edler L, Schmidt R, Hecker E (1982) Program package PAPIL-LOM: computerized acquisition, storage and statistical analysis of mous skin tumor data in chemical co-carcinogenesis, especially tumor promotion. In: Caussinus H et al. (eds) COMPSTAT: Proceedings in computational statistics, 5th symposium held at Toulouse 1982: part II. Short communication, summarized posters. Physika-Verlag, Wien, pp 87–88

    Google Scholar 

  • Gart JJ, Krewski D, Lee PN, Tarone RE, Wahrendorf J (1986) Statistical methods in cancer research, vol III. The design and analysis of long-term animal experiments, IARC Scientific Publications no. 79. International Agency for Research on Cancer, Lyon, pp 69–87

    Google Scholar 

  • Hecker E (1963) über die Wirkstoffe des Crotonöls. I. Biologische Teste zur quantitativen Messung der entzündlichen, cocarcinogenen und toxischen Wirkung. Z Krebsforsch 65:325–333

    Google Scholar 

  • Hecker E (1966) Die cocarcinogene Wirkung der Phorbolester. In: Holzer H, Holldorf AW (Hrsg) 17. Colloquium der Gesellschaft für physiologische Chemie, „Molekularbiologie des malignen Wachstums“. Springer, Berlin Heidelberg New York, S 105–116

    Google Scholar 

  • Hecker E (1967) Phorbol esters from croton oil — chemical nature and biological activities. Naturwissenschaften 54:282–284

    Google Scholar 

  • Hecker E (1968) Cocarcinogenic principles from the seed oil ofCroton tiglium and from other Euphorbiaceae. Cancer Res 28:2338–2349. See also Planta Med [Suppl] (1968) 24–45

    Google Scholar 

  • Hecker E (1971) Isolation and characterization of the co-carcinogenic principles from croton oil. In: Bush H (ed) Methods in cancer research, vol 6. Academic Press, New York, pp 439–484

    Google Scholar 

  • Hecker E (1991a) Wirkungs/Dosis-Beziehungen im Mäusehautmodell der Solitärkanzerogenese mit dem Tumorinitiator 7,12-Dimethylbenz[a]anthrazen als Umweltrisikofaktor — Hypothesen und praktische Konsequenzen. Proceedings 193. Seminar (Workshop) Umweltbundesamt (UBA) „Chemische Krebsrisikofaktoren der Umwelt — Mechanismen ihrer Wirkung und Testsysteme für ihre Erfassung“, Berlin, 23–24 November 1989. Erich-Schmidt-Verlag, Berlin

    Google Scholar 

  • Hecker E (1991b) Wirkungs/Dosis-Beziehungen im Mäusehautmodell der Solitärkanzerogenese mit dem Tumorpromotor 3-O-Tetradekanoylingenol als Umweltrisikofaktor — Hypothesen und praktische Konsequenzen. Proceedings 193. Seminar (Workshop) Umweltbundesamt (UBA) „Chemische Krebsrisikofaktoren der Umwelt — Mechanismen ihrer Wirkung und Testsysteme für ihre Erfassung“, Berlin, 23–24 November 1989. Erich-Schmidt-Verlag, Berlin

    Google Scholar 

  • Hecker E, Rippmann F (1988) Quantitative determination of experimental threshold doses (“no-effect-levels”) for environmental promoters in the initiation/promotion protocol on skin of NMRI-mice. Naunyn-Schmiedeberg's Arch Pharmacol 338 [Suppl]:R11

    Google Scholar 

  • Hecker E, Rippmann F (1989) Assessment of environmental risk factors of Cancer — are there indeed threshold doses? In: Stamatiadis-Smidt H (ed) Deutsches Krebsforschungszentrum, Current Cancer Research 1989. Steinkopff, Darmstadt (Springer, New York), pp 93–103

    Google Scholar 

  • Hecker E, Rippmann F (1990) Outline of a descriptive general theory of environmental chemical cancerogenesis — experimental threshold doses for tumor promoters mechanisms of environmental mutagenesis carcinogenesis. Proceedings of the conference “Mechansisms of environmental mutagenesis-carcinogenesis” 21–26 October 1989, Rhodos, Griechenland. Plenum, New York, pp 167–173

    Google Scholar 

  • Hecker E, Schmidt R (1974) Phorbol esters — the irritants and cocarcinogens ofCroton tiglium L. Prog Chem Org Nat Prod 31:377–467

    Google Scholar 

  • Hergenhahn M, Fürstenberger G, Opferkuch HJ, Adolf W, Mack H, Hecker E (1982) Biological assays for irritant, tumor-initiating and -promoting activities. I. Kinetics of the irritant response in relation to initiation-promoting activity of polyfunctional diterpenes representing tigliane and some daphnane types. J Cancer Res Clin Oncol 104:31–39

    Google Scholar 

  • Hollander M, Wolfe DA (1973) Nonparametric statistical methods. Wiley, New York

    Google Scholar 

  • Iverson K (1987) A dictionary of APL. APL Quote Quad 18:5–40

    Google Scholar 

  • Kalbfleisch JD, Prentice RL (1980) The statistical analysis of failure time data. Wiley, New York

    Google Scholar 

  • Koziol JA, Maxwell DA, Fukushima MC, Colmerauer MEM, Pilch YH (1981) A distribution-free test for tumor growth curve analyses with application to an animal tumor immunotherapy experiment. Biometrics 37:383–390

    Google Scholar 

  • Lotter H, Hecker E (1985) Wechselwirkung von Tumorpromotoren mit zellulären Rezeptoren — feinstruktureller Vergleich von drei Prototypen irritierender und co-carcinogener Diterpenester aus Euphorbiaceen und Thymelaeaceen. Fresenius Z Anal Chem 321:639–640

    Google Scholar 

  • Lotter H, Rippmann F, Hecker E (1991) Structure/activity of polyfunctional diterpenes of the daphnane type. II. Computergraphs of mezerein and phorbol and fine structural comparison of daphnane and tigliane type skin irritants and tumor promoters preparation)

  • Musy C, Morgan KT, Coggins CRE (1981) Computerization of data capture for mouse skin painting studies. Int J Bio-Med Comput 12:419–431

    Google Scholar 

  • Peto R, Pike MC, Day NE, Gray RG, Lee PN, Parish S, Peto J, Richards S, Wahrendorf J (1980) Annex: guidelines for simple, sensitive significance tests for carcinogenic effects in long-term animal experiments. IARC monographs on the evaluation of the carcinogenic risk of chemicals to humans [Suppl 2]. In: Long-term and short-term screening assays for carcinogens: a critical appraisal. International Agency for Research on Cancer, Lyon, pp 311–426

    Google Scholar 

  • Rippmann F (1987) Quantifizierung und Differenzierung der solitärkarzinogenen Wirkung von DMBA und der tumorpromovierenden Wirkung von TPA, 3-TI und Simplexin im Modell der Mäusehaut. Dissertation, Universität Heidelberg

  • Rippmann F, Hecker E (1988) General quantitative dose/time(response) relationship for solitary cancerogenesis and initiation/ promotion in skin of NMRI-mice and its implications for risk assessment. Naunyn-Schmiedeberg's Arch Pharmacol 338 [Suppl]:R86

    Google Scholar 

  • Rippmann F (1990) Hydrophobicity and tumor promoting activity of phorbol esters. Quant Struct -Act Relat 9:1–5

    Google Scholar 

  • SAS Institute Inc (1985) SAS user's guide: statistics, 5th edn. SAS Institute Inc, Cary NC

    Google Scholar 

  • Schmidt R, Hecker E (1989) Biological assays for irritant, tumor initiating and -promoting activities. II. Standardized initiation/promotion protocol and semiquantitative estimation of promoting (or initiating) potencies in skin of NMRI mice. J Cancer Res Clin Oncol 115:516–524

    Google Scholar 

  • Weber E (ed) (1980) ADAM — Statistische Auswertung Biomedizinischer Daten, Teil: 1. Datenerfassungs- und Auswertungssystem. Internal publication, DKFZ, Heidelberg

    Google Scholar 

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Authors and Affiliations

  1. Institute of Epidemiology and Biometry, Im Neuenheimer Feld 280, D-6900, Heidelberg, Federal Republic of Germany

    Lutz Edler & Ernst Weber

  2. German Cancer Research Center, Institute of Biochemistry, Im Neuenheimer Feld 280, D-6900, Heidelberg, Federal Republic of Germany

    Rainer Schmidt, Friedrich Rippmann & Erich Hecker

Authors
  1. Lutz Edler
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  2. Rainer Schmidt
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  3. Ernst Weber
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  4. Friedrich Rippmann
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  5. Erich Hecker
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Additional information

Dedicated to Prof. I. Bernhard Weinstein, Director Comprehensive Cancer Center, College of Physicians and Surgeons, Columbia University, New York, USA, on the occasion of his 60th birthday in appreciation of his fascinating contributions to many aspects of tumor promotion.

For paper II in this series see Schmidt and Hecker (1989)

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Edler, L., Schmidt, R., Weber, E. et al. Biological assays for irritant, tumor-initiating and -promoting activities. J Cancer Res Clin Oncol 117, 205–216 (1991). https://doi.org/10.1007/BF01625426

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  • DOI: https://doi.org/10.1007/BF01625426

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